Synergistic water soluble preservative compositions of biocidal mixtures

ABSTRACT

A water soluble preservative antimicrobial composition for addition to commercial use products at predetermined use levels to provide synergistic biocidal activity against a wide range of fungi and gram-negative and gram-positive bacteria, comprising an active liquid blend of 
     (1) an admixture of 
     (a) one or more methylol compounds selected from the group consisting of N-[1,3-bis (hydroxymethyl)-2,5-dioxo-4-imidazolidinyl]-N,N&#39;-bis (hydroxymethyl) urea; 1,3-dimethylol-5,5-dimethyl hydantoin; sodium hydroxymethyl glycinate; 1,1-methylene bis[3-[3-(hydroxymethyl)-2,5-dioxo-4-imidazolidinyl]urea]; and glycine anhydride dimethylol; and 
     (b) 3-iodo-2-propynylbutyl carbamate, in a weight ratio of (a):(b) of 100:1 to 2000:1, and 
     (2) propylene glycol or 1,3-butylene glycol.

CROSS-REFERENCE TO RELATED PATENT APPLICATION

This is a division of application Ser. No. 08/413,742, filed Mar. 30, 1995, now U.S. Pat. No. 5,552,425, which is a continuation-in-part of Ser. No. 08/234,089, filed Apr. 28, 1994 now U.S. Pat. No. 5,428,050.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to preservative compositions, and, more particularly, to a water soluble antibacterial compositions for addition to commercial use products to provide long time synergistic biocidal activity therein, which are active liquid blends of an admixture of a methylol compound and an iodopropynyl compound, in a predetermined weight ratio of 100:1 to 2000:1, and propylene glycol or 1,3-butylene glycol.

2. Description of the Prior Art

Combinations of antimicrobial agents have been developed in the prior art in order to:

(1) produce a biochemical synergism;

(2) broaden the antimicrobial spectrum of activity of each agent;

(3) increase water solubility for the admixture;

(4) minimize the toxicity or irritation of a given agent to the host; and

(5) minimize physical and chemical incompatibilities.

True biological synergism exists when two agents, when combined, require lesser amounts of the agents to bring about the same inhibitory or cidal effect than either single agent alone. While synergistic interaction for two or more antimicrobial agents does produce more than merely an additive effect in the resultant biological activity, in most cases the mechanism of such synergism remains a mystery.

M. Rosen et al., in U.S. Pat. No. 4,844,891, for example, described a preservative admixture of (a) a formaldehyde donor and (b) a halopropynyl compound, in a weight ratio of (a):(b) of 50:1 to 1:1, preferably 30:1 to 2:1, and, most preferably, 20:1 to 10:1, as providing fungicidal activity for 1-3 days in commercial use formulations. However, Rosen observed that when the ratio of (a):(b) in the concentrate exceeded 50:1 (System No. 16 in Table 1, a ratio of 73.33), the preservative composition was ineffective in providing biocidal protection in the use formulations. Thus a relatively large amount of the halopropynyl compound was required by Rosen to provide significant biocidal activity in the use composition. In such admixtures, although the formaldehyde donor is water soluble, the halopropynyl compound is substantially insoluble in water. Therefore it was difficult for Rosen to uniformly distribute his admixture throughout the use composition.

For these and other reasons, it is desired to provide a new and improved water soluble preservative admixture of such biocidal compounds which requires relatively little of the water insoluble and expensive halopropynyl compound, and that also provides effective synergistic protection in use formulations against a wide range of fungi and bacteria at different use levels over a long period of time.

A feature of the present invention is the provision of an admixture concentrate which is water soluble and which therefore can be uniformly distributed in use compositions at a predetermined use level.

Another feature herein is the provision of a preservative admixture which exhibits a long term synergistic biocidal activity against wide range of fungi and both gram-negative and gram-positive bacteria in the use compositions.

Still another feature of this invention is the provision of a water soluble preservative antibacterial composition for addition to commercial use products at predetermined use levels against a wide range of fungi and gram-negative and gram-positive bacteria, in the form of an active liquid blend of an admixture of one or more methylol compounds and an iodopropynyl compound, in a predetermined weight ratio of 100:1 to 2000:1, and propylene glycol or 1,3-butylene glycol.

These and other objects and features of the invention will be made apparent from the following more particular description of the invention.

SUMMARY OF THE INVENTION

What has been discovered is a water soluble preservative composition for addition to commercial use compositions at predetermined use levels, and uniformly distributed therein, which provides long term synergistic biocidal activity against a wide range of fungi and both gram-negative and gram-positive bacteria. The composition of the invention comprises:

(1) an admixture of powders of

(a) a methylol compound, or their equivalent, and

(b) iodopropynyl alcohol, or its ester, carbamate or ether derivative thereof,

in a weight ratio of (a):(b) of 100:1 to 2000:1, preferably 200:1 to 500:1; and

(2) propylene glycol or 1,3-butylene glycol.

Suitably, the composition of the invention comprises about 20 to 55 weight percent of (1) and 45-80 weight percent of (2), preferably 40 weight percent of (1) and 60 weight percent of (2).

Commercial use compositions containing about 0.1 to 5% by weight, preferably 0.5-1%, of the composition of the invention also are provided therein. Such use compositions contain an iodopropynyl compound in an amount necessary to provide the desired antifungal activity, and a methylol compound in an amount necessary to provide the desired antibacterial activity for the user.

DETAILED DESCRIPTION OF THE INVENTION

The invention is based upon the discoveries that in a predetermined admixture of (a) a methylol compound, or its equivalent, and (b) an iodopropynyl compound:

(1) Iodopropynyl compounds are substantially water insoluble at weight ratios of (a):(b) of less than 100; accordingly, at weight ratios below 100:1, it is difficult to uniformly distribute the iodopropynyl compound in aqueous use compositions, particularly in creams, gels and the like. In this invention, the admixtures are used at a weight ratio of (a):(b) of 100:1 to 2000:1, which are water soluble in all use compositions at conventional use levels.

(2) Effective synergistic biocidal activity is achieved for admixtures having a weight ratio of (a):(b) of 100:1 to 2000:1. Such admixtures have a Synergistic Index (SI) value approaching zero (maximum synergism) for a wide range of organisms. In contrast, admixtures with (a):(b) ratios below 100:1, e.g. 10:1 to 50:1, are much less synergistic, and are active with only a narrower range of organisms.

(3) A water soluble preservative antibacterial composition having enhanced activity can be made available in the form of a liquid blend of an admixture of one or more methyol compounds and 3-iodo-2-propynylbutyl carbamate, in a weight ratio of 100:1 to 2000:1, and propylene glycol or 1,3-butylene glycol, in a predetermined formulation of 20-55 weight percent of the admixture and 45-80 weight percent of the glycol, preferably 40:60.

(4) Preservative activity for use compositions is achieved most effectively with an admixture wt. ratio of 100:1 to 2000:1 at use levels of the composition of 0.1 to 5%, preferably 0.5 to 1%, by weight of the finished product. In this amount, the antifungal iodopropynyl compound is present in a small amount which provides antifungal protection and which significantly reduces the cost and toxicity of the use composition, and the antibacterial methylol compound is present in an amount which is sufficient to provide the desired antibacterial protection.

The experimental results upon which these discoveries are based are described below. In these examples, the (a) methylol compound may be selected from diazolidinyl urea (GERMALL® II) N-[1,3-bis(hydroxymethyl)-2,5-dioxo-4-imidazolidinyl]-N,N'-bis(hydroxymethyl) urea, imidurea (GERMALL® 115), 1,1-methylene bis[3-[3-(hydroxymethyl)-2,5-dioxo-4-imidazolidinyl]urea]; 1,3-dimethylol-5,5-dimethyl hydantoin (DMDMH), sodium hydroxymethyl glycinate (SUTTOCIDE® A), and glycine anhydride dimethylol (GADM); the (b) iodopropynyl compound is 3-iodo-2-propynylbutyl carbamate (IPBC); and the (c) glycol is propylene glycol or 1,3-butylene glycol.

1. WATER SOLUBILITY

The water solubility or insolubility of admixtures of several methylol compounds with IPBC as 1% aqueous solutions is shown in Tables A through C below.

                  TABLE A                                                          ______________________________________                                         Weight Ratio of                                                                Germall ® II:IPBC                                                                          Amount in ppm                                                                              Solubility                                         ______________________________________                                         2000:1           5          Soluble                                            1000:1          10          Soluble                                            500:1           20          Soluble                                            200:1           50          Soluble                                            100:1           100         Soluble                                             50:1           200         Insoluble                                           20:1           500         Insoluble                                          ______________________________________                                    

                  TABLE B                                                          ______________________________________                                         Weight Ratio of                                                                GADM:IPBC      Amount in ppm                                                                              Solubility                                          ______________________________________                                         2000:1          5          Soluble                                             1000:1         10          Soluble                                             500:1          20          Soluble                                             200:1          50          Soluble                                             100:1          100         Soluble                                              50:1          200         Insoluble                                            20:1          500         Insoluble                                           ______________________________________                                    

                  TABLE C                                                          ______________________________________                                         Weight Ratio of                                                                DMDMH:IPBC     Amount in ppm                                                                              Solubility                                          ______________________________________                                         2000:1          5          Soluble                                             1000:1         10          Soluble                                             500:1          20          Soluble                                             200:1          50          Soluble                                             100:1          100         Soluble                                              50:1          200         Insoluble                                            20:1          500         Insoluble                                           ______________________________________                                    

These results demonstrate that admixtures having a ratio of 50:1 or 20:1 are insoluble in water whereas at ratios of 100:1 to 2000:1 the admixtures are soluble in water.

2. SYNERGISM

Tables 1 through 14 below demonstrate the very effective synergistic interaction between compounds "a" and "b" against a broad range of fungi and both gram-negative and gram-positive bacteria. The following organisms were tested:

    ______________________________________                                                      ATCC                                                              Organism     Number   Inoculum Concentration                                   ______________________________________                                         Ps. aeruginosa (PSA)*                                                                        9027    2.1 × 10.sup.6 CFU/gm of Product                   E. coli (ECOLI)*                                                                             8739    4.7 × 10.sup.5 CFU/gm of Product                   Staph. aureus (SA)**                                                                         6538    1.6 × 10.sup.6 CFU/gm of Product                   Ps. cepacia (PC)*                                                                           25416    1.6 × 10.sup.6 CFU/gm of Product                   C. albicans (CAN)***                                                                        10231    8.0 × 10.sup.4 CFU/gm of Product                   A. niger (AN)***                                                                            16404    2.7 × 10.sup.5 CFU/gm of Product                   ______________________________________                                          *gram-negative bacteria                                                        **grampositive bacteria                                                        ***fungi                                                                 

Table D below lists the static (MIC) and cidal activities of several antimicrobial agents in ppm. These activities are used to calculate the Synergism Index (SI) of admixtures of the present invention.

                                      TABLE D                                      __________________________________________________________________________     Static (MIC) and Cidal Activities of Several Antimicrobial Compounds           (Static/Cidal Concentrations in ppm)                                           Organism                                                                       (ATCC #) IPBC  Germall ® II                                                                      GADM   DMDMH  IPGA                                       __________________________________________________________________________     (SA) (6538)                                                                             100/200                                                                               400/1600                                                                             400/800                                                                                450/1600                                                                              300/5000                                  (ECOLI) (8739)                                                                           50/100                                                                               400/1600                                                                             400/800                                                                               400/800                                                                               150/600                                    (PSA) (9027)                                                                             800/1200                                                                             400/1600                                                                             400/400                                                                                600/1600                                                                             70/70                                      (PC) (25416)                                                                            1200/1800                                                                            200/400                                                                               200/400                                                                                600/1600                                                                              70/300                                    (CAN) (10231)                                                                            50/100                                                                               1500/15000                                                                            7500/15000                                                                            8000/16000                                                                            50/300                                    (AN) (16404)                                                                             50/100                                                                              3200/3200                                                                             1600/3200                                                                              8000/16000                                                                           30/30                                      __________________________________________________________________________

The Synergism Index was determined by the same mathematical treatment of such data described by Kull et al. in Applied Microbiology 9, 538-541 (1961) using the following relationship: ##EQU1## where: 1. Q_(a) =The quantity of Compound a acting alone, producing an endpoint.

2. Q_(b) =The quantity of Compound b acting alone, producing and endpoint.

3. Q_(A) =The quantity of Compound A in mixture, producing an endpoint.

4. Q_(B) =The quantity of Compound B in mixture, producing an endpoint.

When SI is equal to 1, a mere additive effect of the components in the mixture is indicated; when SI is less than 1, synergism has occurred; and when SI is greater than 1 it indicates antagonism of the two components.

According to this well known method of measuring synergism, the quantity of each component in the various mixtures is compared with the quantity of pure component that is required to reach the same endpoint or to produce the same microbiological effect as the mixture.

                  TABLE 1                                                          ______________________________________                                         2000:1 Wt. Ratio GERMALL ® II/IPBC                                         Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  0.05 99.95  0.06                               "        ECOLI    100     1600  0.05 99.95  0.06                               "        PSA      1200    1600  0.05 99.95  0.06                               "        PC       1800    1250  0.05 99.95  0.08                               "        CAN      100     15000 0.05 99.95  0.01                               "        AN       100     3200  0.05 99.95  0.03                                0.025%  SA       200     1600  0.125                                                                               249.9  0.16                               "        ECOLI    100     1600  0.125                                                                               249.9  0.16                               "        PSA      1200    1600  0.125                                                                               249.9  0.16                               "        PC       1800    1250  0.125                                                                               249.9  0.20                               "        CAN      100     15000 0.125                                                                               249.9  0.02                               "        AN       100     3200  0.125                                                                               249.9  0.08                               0.05%    SA       200     1600  0.25 499.8  0.31                               "        ECOLI    100     1600  0.25 499.8  0.31                               "        PSA      1200    1600  0.25 499.8  0.31                               "        PC       1800    1250  0.25 499.8  0.40                               "        CAN      100     15000 0.25 499.8  0.04                               "        AN       100     3200  0.25 499.8  0.16                               0.10%    SA       200     1600  0.5  999.5  0.63                               "        ECOLI    100     1600  0.5  999.5  0.63                               "        PSA      1200    1600  0.5  999.5  0.63                               "        PC       1800    1250  0.5  999.5  0.80                               "        CAN      100     15000 0.5  999.5  0.07                               "        AN       100     3200  0.5  999.5  0.32                               0.20%    SA       200     1600  1    1999   1.25                               "        ECOLI    100     1600  1    1999   1.26                               "        PSA      1200    1600  1    1999   1.25                               "        PC       1800    1250  1    1999   1.60                               "        CAN      100     15000 1    1999   0.14                               "        AN       100     3200  1    1999   0.63                               0.40%    SA       200     1600  2    3998   2.51                               "        ECOLI    100     1600  2    3998   2.52                               "        PSA      1200    1600  2    3998   2.50                               "        PC       1800    1250  2    3998   3.20                               "        CAN      100     15000 2    3998   0.29                               "        AN       100     3200  2    3998   1.27                               0.50%    SA       200     1600  2.5  4997.5 3.14                               "        ECOLI    100     1600  2.5  4997.5 3.15                               "        PSA      1200    1600  2.5  4997.5 3.13                               "        PC       1800    1250  2.5  4997.5 4.00                               "        CAN      100     15000 2.5  4997.5 0.36                               "        AN       100     3200  2.5  4997.5 1.59                               ______________________________________                                    

                  TABLE 2                                                          ______________________________________                                         1000:1 Wt. Ratio GERMALL ® II/IPBC                                         Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  0.1  99.9   0.06                               "        ECOLI    100     1600  0.1  99.9   0.06                               "        PSA      1200    1600  0.1  99.9   0.06                               "        PC       1800    1250  0.1  99.9   0.08                               "        CAN      100     15000 0.1  99.9   0.01                               "        AN       100     3200  0.1  99.9   0.03                                0.025%  SA       200     1600  0.25 249.8  0.16                               "        ECOLI    100     1600  0.25 249.8  0.16                               "        PSA      1200    1600  0.25 249.8  0.16                               "        PC       1800    1250  0.25 249.8  0.20                               "        CAN      100     15000 0.25 249.8  0.02                               "        AN       100     3200  0.25 249.8  0.08                               0.05%    SA       200     1600  0.5  499.5  0.31                               "        ECOLI    100     1600  0.5  499.5  0.32                               "        PSA      1200    1600  0.5  499.5  0.31                               "        PC       1800    1250  0.5  499.5  0.40                               "        CAN      100     15000 0.5  499.5  0.04                               "        AN       100     3200  0.5  499.5  0.16                               0.10%    SA       200     1600  1    999    0.63                               "        ECOLI    100     1600  1    999    0.63                               "        PSA      1200    1600  1    999    0.63                               "        PC       1800    1250  1    999    0.80                               "        CAN      100     15000 1    999    0.08                               "        AN       100     3200  1    999    0.32                               0.20%    SA       200     1600  2    1998   1.26                               "        ECOLI    100     1600  2    1998   1.27                               "        PSA      1200    1600  2    1998   1.25                               "        PC       1800    1250  2    1998   1.60                               "        CAN      100     15000 2    1998   0.15                               "        AN       100     3200  2    1998   0.64                               0.40%    SA       200     1600  4    3996   2.52                               "        ECOLI    100     1600  4    3996   2.54                               "        PSA      1200    1600  4    3996   2.50                               "        PC       1800    1250  4    3996   3.20                               "        CAN      100     15000 4    3996   0.31                               "        AN       100     3200  4    3996   1.29                               0.50%    SA       200     1600  5    4995   3.15                               "        ECOLI    100     1600  5    4995   3.17                               "        PSA      1200    1600  5    4995   3.13                               "        PC       1800    1250  5    4995   4.00                               "        CAN      100     15000 5    4995   0.38                               "        AN       100     3200  5    4995   1.61                               ______________________________________                                    

                  TABLE 3                                                          ______________________________________                                         500:1 Wt. Ratio GERMALL ® II/IPBC                                          Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  0.2  99.8   0.06                               "        ECOLI    100     1600  0.2  99.8   0.06                               "        PSA      1200    1600  0.2  99.8   0.06                               "        PC       1800    1250  0.2  99.8   0.08                               "        CAN      100     15000 0.2  99.8   0.01                               "        AN       100     3200  0.2  99.8   0.03                                0.025%  SA       200     1600  0.5  249.5  0.16                               "        ECOLI    100     1600  0.5  249.5  0.16                               "        PSA      1200    1600  0.5  249.5  0.16                               "        PC       1800    1250  0.5  249.5  0.20                               "        CAN      100     15000 0.5  249.5  0.02                               "        AN       100     3200  0.5  249.5  0.08                               0.05%    SA       200     1600  1    499    0.32                               "        ECOLI    100     1600  1    499    0.32                               "        PSA      1200    1600  1    499    0.31                               "        PC       1800    1250  1    499    0.40                               "        CAN      100     15000 1    499    0.04                               "        AN       100     3200  1    499    0.17                               0.10%    SA       200     1600  2    998    0.63                               "        ECOLI    100     1600  2    998    0.64                               "        PSA      1200    1600  2    998    0.63                               "        PC       1800    1250  2    998    0.80                               "        CAN      100     15000 2    998    0.09                               "        AN       100     3200  2    998    0.33                               0.20%    SA       200     1600  4    1996   1.27                               "        ECOLI    100     1600  4    1996   1.29                               "        PSA      1200    1600  4    1996   1.25                               "        PC       1800    1250  4    1996   1.60                               "        CAN      100     15000 4    1996   0.17                               "        AN       100     3200  4    1996   0.66                               0.40%    SA       200     1600  8    3992   2.54                               "        ECOLI    100     1600  8    3992   2.58                               "        PSA      1200    1600  8    3992   2.50                               "        PC       1800    1250  8    3992   3.20                               "        CAN      100     15000 8    3992   0.35                               "        AN       100     3200  8    3992   1.33                               0.50%    SA       200     1600  10   4990   3.17                               "        ECOLI    100     1600  10   4990   3.22                               "        PSA      1200    1600  10   4990   3.13                               "        PC       1800    1250  10   4990   4.00                               "        CAN      100     15000 10   4990   0.43                               "        AN       100     3200  10   4990   1.66                               ______________________________________                                    

                  TABLE 4                                                          ______________________________________                                         200:1 Wt. Ratio GERMALL ® II/IPBC                                          Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  0.5  99.5   0.06                               "        ECOLI    100     1600  0.5  99.5   0.07                               "        PSA      1200    1600  0.5  99.5   0.06                               "        PC       1800    1250  0.5  99.5   0.08                               "        CAN      100     15000 0.5  99.5   0.01                               "        AN       100     3200  0.5  99.5   0.04                                0.025%  SA       200     1600  1.25 248.75 0.16                               "        ECOLI    100     1600  1.25 248.75 0.17                               "        PSA      1200    1600  1.25 248.75 0.16                               "        PC       1800    1250  1.25 248.75 0.20                               "        CAN      100     15000 1.25 248.75 0.03                               "        AN       100     3200  1.25 248.75 0.09                               0.05%    SA       200     1600  2.5  497.5  0.32                               "        ECOLI    100     1600  2.5  497.5  0.34                               "        PSA      1200    1600  2.5  497.5  0.31                               "        PC       1800    1250  2.5  497.5  0.40                               "        CAN      100     15000 2.5  497.5  0.06                               "        AN       100     3200  2.5  497.5  0.18                               0.10%    SA       200     1600  5    995    0.65                               "        ECOLI    100     1600  5    995    0.67                               "        PSA      1200    1600  5    995    0.63                               "        PC       1800    1250  5    995    0.80                               "        CAN      100     15000 5    995    0.12                               "        AN       100     3200  5    995    0.36                               0.20%    SA       200     1600  10   1990   1.29                               "        ECOLI    100     1600  10   1990   1.34                               "        PSA      1200    1600  10   1990   1.25                               "        PC       1800    1250  10   1990   1.60                               "        CAN      100     15000 10   1990   0.23                               "        AN       100     3200  10   1990   0.72                               0.40%    SA       200     1600  20   3980   2.59                               "        ECOLI    100     1600  20   3980   2.69                               "        PSA      1200    1600  20   3980   2.50                               "        PC       1800    1250  20   3980   3.20                               "        CAN      100     15000 20   3980   0.47                               "        AN       100     3200  20   3980   1.44                               0.50%    SA       200     1600  25   4975   3.23                               "        ECOLI    100     1600  25   4975   3.36                               "        PSA      1200    1600  25   4975   3.13                               "        PC       1800    1250  25   4975   3.99                               "        CAN      100     15000 25   4975   0.58                               "        AN       100     3200  25   4975   1.80                               ______________________________________                                    

                  TABLE 5                                                          ______________________________________                                         100:1 Wt. Ratio GERMALL ® II/IPBC                                          Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  1    99     0.07                               "        ECOLI    100     1600  1    99     0.07                               "        PSA      1200    1600  1    99     0.06                               "        PC       1800    1250  1    99     0.08                               "        CAN      100     15000 1    99     0.02                               "        AN       100     3200  1    99     0.04                                0.025%  SA       200     1600  2.5  248    0.17                               "        ECOLI    100     1600  2.5  248    0.18                               "        PSA      1200    1600  2.5  248    0.16                               "        PC       1800    1250  2.5  248    0.20                               "        CAN      100     15000 2.5  248    0.04                               "        AN       100     3200  2.5  248    0.10                               0.05%    SA       200     1600  5    495    0.33                               "        ECOLI    100     1600  5    495    0.36                               "        PSA      1200    1600  5    495    0.31                               "        PC       1800    1250  5    495    0.40                               "        CAN      100     15000 5    495    0.08                               "        AN       100     3200  5    495    0.20                               0.10%    SA       200     1600  10   990    0.67                               "        ECOLI    100     1600  10   990    0.72                               "        PSA      1200    1600  10   990    0.63                               "        PC       1800    1250  10   990    0.80                               "        CAN      100     15000 10   990    0.17                               "        AN       100     3200  10   990    0.41                               0.20%    SA       200     1600  20   1980   1.34                               "        ECOLI    100     1600  20   1980   1.44                               "        PSA      1200    1600  20   1980   1.25                               "        PC       1800    1250  20   1980   1.60                               "        CAN      100     15000 20   1980   0.33                               "        AN       100     3200  20   1980   0.82                               0.40%    SA       200     1600  40   3960   2.68                               "        ECOLI    100     1600  40   3960   2.88                               "        PSA      1200    1600  40   3960   2.51                               "        PC       1800    1250  40   3960   3.19                               "        CAN      100     15000 40   3960   0.66                               "        AN       100     3200  40   3960   1.64                               0.50%    SA       200     1600  50   4950   3.34                               "        ECOLI    100     1600  50   4950   3.59                               "        PSA      1200    1600  50   4950   3.14                               "        PC       1800    1250  50   4950   3.99                               "        CAN      100     15000 50   4950   0.83                               "        AN       100     3200  50   4950   2.05                               ______________________________________                                    

                  TABLE 6                                                          ______________________________________                                         50:1 Wt. Ratio GERMALL ® II/IPBC                                           Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  2    98     0.07                               "        ECOLI    100     1600  2    98     0.08                               "        PSA      1200    1600  2    98     0.06                               "        PC       1800    1250  2    98     0.08                               "        CAN      100     15000 2    98     0.03                               "        AN       100     3200  2    98     0.05                                0.025%  SA       200     1600  5    245    0.18                               "        ECOLI    100     1600  5    245    0.20                               "        PSA      1200    1600  5    245    0.16                               "        PC       1800    1250  5    245    0.20                               "        CAN      100     15000 5    245    0.07                               "        AN       100     3200  5    245    0.13                               0.05%    SA       200     1600  10   490    0.36                               "        ECOLI    100     1600  10   490    0.41                               "        PSA      1200    1600  10   490    0.31                               "        PC       1800    1250  10   490    0.40                               "        CAN      100     15000 10   490    0.13                               "        AN       100     3200  10   490    0.25                               0.10%    SA       200     1600  20   980    0.71                               "        ECOLI    100     1600  20   980    0.81                               "        PSA      1200    1600  20   980    0.63                               "        PC       1800    1250  20   980    0.80                               "        CAN      100     15000 20   980    0.27                               "        AN       100     3200  20   980    0.51                               0.20%    SA       200     1600  40   1960   1.43                               "        ECOLI    100     1600  40   1960   1.63                               "        PSA      1200    1600  40   1960   1.26                               "        PC       1800    1250  40   1960   1.59                               "        CAN      100     15000 40   1960   0.53                               "        AN       100     3200  40   1960   1.01                               0.40%    SA       200     1600  80   3920   2.85                               "        ECOLI    100     1600  80   3920   3.25                               "        PSA      1200    1600  80   3920   2.52                               "        PC       1800    1250  80   3920   3.18                               "        CAN      100     15000 80   3920   1.06                               "        AN       100     3200  80   3920   2.03                               0.50%    SA       200     1600  100  4900   3.56                               "        ECOLI    100     1600  100  4900   4.06                               "        PSA      1200    1600  100  4900   3.15                               "        PC       1800    1250  100  4900   3.98                               "        CAN      100     15000 100  4900   1.33                               "        AN       100     3200  100  4900   2.53                               ______________________________________                                    

                  TABLE 7                                                          ______________________________________                                         20:1 Wt. Ratio GERMALL ® II/IPBC                                           Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  5    95     0.08                               "        ECOLI    100     1600  5    95     0.11                               "        PSA      1200    1600  5    95     0.06                               "        PC       1800    1250  5    95     0.08                               "        CAN      100     15000 5    95     0.06                               "        AN       100     3200  5    95     0.08                                0.025%  SA       200     1600  12.5 237.5  0.21                               "        ECOLI    100     1600  12.5 237.5  0.27                               "        PSA      1200    1600  12.5 237.5  0.16                               "        PC       1800    1250  12.5 237.5  0.20                               "        CAN      100     15000 12.5 237.5  0.14                               "        AN       100     3200  12.5 237.5  0.20                               0.05%    SA       200     1600  25   475    0.42                               "        ECOLI    100     1600  25   475    0.55                               "        PSA      1200    1600  25   475    0.32                               "        PC       1800    1250  25   475    0.39                               "        CAN      100     15000 25   475    0.28                               "        AN       100     3200  25   475    0.40                               0.10%    SA       200     1600  50   950    0.84                               "        ECOLI    100     1600  50   950    1.09                               "        PSA      1200    1600  50   950    0.64                               "        PC       1800    1250  50   950    0.79                               "        CAN      100     15000 50   950    0.56                               "        AN       100     3200  50   950    0.80                               0.20%    SA       200     1600  100  1900   1.69                               "        ECOLI    100     1600  100  1900   2.19                               "        PSA      1200    1600  100  1900   1.27                               "        PC       1800    1250  100  1900   1.58                               "        CAN      100     15000 100  1900   1.13                               "        AN       100     3200  100  1900   1.59                               0.40%    SA       200     1600  200  4800   4.00                               "        ECOLI    100     1600  200  4800   5.00                               "        PSA      1200    1600  200  4800   3.17                               "        PC       1800    1250  200  4800   3.95                               "        CAN      100     15000 200  4800   2.32                               "        AN       100     3200  200  4800   3.50                               0.50%    SA       200     1600  250  4750   4.22                               "        ECOLI    100     1600  250  4750   5.47                               "        PSA      1200    1600  250  4750   3.18                               "        PC       1800    1250  250  4750   3.94                               "        CAN      100     15000 250  4750   2.82                               "        AN       100     3200  250  4750   3.98                               ______________________________________                                    

                  TABLE 8                                                          ______________________________________                                         2000:1 Wt. Ratio DMDMH/IPBC                                                    Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  0.05 99.95  0.06                               "        ECOLI    100     800   0.05 99.95  0.13                               "        PSA      1200    1600  0.05 99.95  0.06                               "        PC       1800    1600  0.05 99.95  0.06                               "        CAN      100     16000 0.05 99.95  0.01                               "        AN       100     16000 0.05 99.95  0.01                                0.025%  SA       200     1600  0.125                                                                               249.88 0.16                               "        ECOLI    100     800   0.125                                                                               249.88 0.31                               "        PSA      1200    1600  0.125                                                                               249.88 0.16                               "        PC       1800    1600  0.125                                                                               249.88 0.16                               "        CAN      100     16000 0.125                                                                               249.88 0.02                               "        AN       100     16000 0.125                                                                               249.88 0.02                               0.05%    SA       200     1600  0.25 499.75 0.31                               "        ECOLI    100     800   0.25 499.75 0.63                               "        PSA      1200    1600  0.25 499.75 0.31                               "        PC       1800    1600  0.25 499.75 0.31                               "        CAN      100     16000 0.25 499.75 0.03                               "        AN       100     16000 0.25 499.75 0.03                               0.10%    SA       200     1600  0.5  999.5  0.63                               "        ECOLI    100     800   0.5  999.5  1.25                               "        PSA      1200    1600  0.5  999.5  0.63                               "        PC       1800    1600  0.5  999.5  0.62                               "        CAN      100     16000 0.5  999.5  0.07                               "        AN       100     16000 0.5  999.5  0.07                               0.20%    SA       200     1600  1    1999   1.25                               "        ECOLI    100     800   1    1999   2.51                               "        PSA      1200    1600  1    1999   1.25                               "        PC       1800    1600  1    1999   1.25                               "        CAN      100     16000 1    1999   0.13                               "        AN       100     16000 1    1999   0.13                               0.40%    SA       200     1600  2    3998   2.51                               "        ECOLI    100     800   2    3998   5.02                               "        PSA      1200    1600  2    3998   2.50                               "        PC       1800    1600  2    3998   2.50                               "        CAN      100     16000 2    3998   0.27                               "        AN       100     16000 2    3998   0.27                               0.50%    SA       200     1600  2.5  4997.5 3.14                               "        ECOLI    100     800   2.5  4997.5 6.27                               "        PSA      1200    1600  2.5  4997.5 3.13                               "        PC       1800    1600  2.5  4997.5 3.12                               "        CAN      100     16000 2.5  4997.5 0.34                               "        AN       100     16000 2.5  4997.5 0.34                               ______________________________________                                    

                  TABLE 9                                                          ______________________________________                                         1000:1 Wt. Ratio DMDMH/IPBC                                                    Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  0.1  99.9   0.06                               "        ECOLI    100     800   0.1  99.9   0.13                               "        PSA      1200    1600  0.1  99.9   0.06                               "        PC       1800    1600  0.1  99.9   0.06                               "        CAN      100     16000 0.1  99.9   0.01                               "        AN       100     16000 0.1  99.9   0.01                                0.025%  SA       200     1600  0.25 249.8  0.16                               "        ECOLI    100     800   0.25 249.8  0.31                               "        PSA      1200    1600  0.25 249.8  0.16                               "        PC       1800    1600  0.25 249.8  0.16                               "        CAN      100     16000 0.25 249.8  0.02                               "        AN       100     16000 0.25 249.8  0.02                               0.05%    SA       200     1600  0.5  499.5  0.31                               "        ECOLI    100     800   0.5  499.5  0.63                               "        PSA      1200    1600  0.5  499.5  0.31                               "        PC       1800    1600  0.5  499.5  0.31                               "        CAN      100     16000 0.5  499.5  0.04                               "        AN       100     16000 0.5  499.5  0.04                               0.10%    SA       200     1600  1    999    0.63                               "        ECOLI    100     800   1    999    1.26                               "        PSA      1200    1600  1    999    0.63                               "        PC       1800    1600  1    999    0.62                               "        CAN      100     16000 1    999    0.07                               "        AN       100     16000 1    999    0.07                               0.20%    SA       200     1600  2    1998   1.26                               "        ECOLI    100     800   2    1998   2.52                               "        PSA      1200    1600  2    1998   1.25                               "        PC       1800    1600  2    1998   1.25                               "        CAN      100     16000 2    1998   0.14                               "        AN       100     16000 2    1998   0.14                               0.40%    SA       200     1600  4    3996   2.52                               "        ECOLI    100     800   4    3996   5.04                               "        PSA      1200    1600  4    3996   2.50                               "        PC       1800    1600  4    3996   2.50                               "        CAN      100     16000 4    3996   0.29                               "        AN       100     16000 4    3996   0.29                               0.50%    SA       200     1600  5    4995   3.15                               "        ECOLI    100     800   5    4995   6.29                               "        PSA      1200    1600  5    4995   3.13                               "        PC       1800    1600  5    4995   3.12                               "        CAN      100     16000 5    4995   0.36                               "        AN       100     16000 2.5  4997.5 0.36                               ______________________________________                                    

                  TABLE 10                                                         ______________________________________                                         500:1 Wt. Ratio DMDMH/IPBC                                                     Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  0.2  99.8   0.06                               "        ECOLI    100     800   0.2  99.8   0.13                               "        PSA      1200    1600  0.2  99.8   0.06                               "        PC       1800    1600  0.2  99.8   0.06                               "        CAN      100     16000 0.2  99.8   0.01                               "        AN       100     16000 0.2  99.8   0.01                               0.25%    SA       200     1600  0.5  249.5  0.16                               "        ECOLI    100     800   0.5  249.5  0.32                               "        PSA      1200    1600  0.5  249.5  0.16                               "        PC       1800    1600  0.5  249.5  0.16                               "        CAN      100     16000 0.5  249.5  0.02                               "        AN       100     16000 0.5  249.5  0.02                               0.05%    SA       200     1600  1    499    0.32                               "        ECOLI    100     800   1    499    0.63                               "        PSA      1200    1600  1    499    0.31                               "        PC       1800    1600  1    499    0.31                               "        CAN      100     16000 1    499    0.04                               "        AN       100     16000 1    499    0.04                               0.10%    SA       200     1600  2    998    0.63                               "        ECOLI    100     800   2    998    1.27                               "        PSA      1200    1600  2    998    0.63                               "        PC       1800    1600  2    998    0.62                               "        CAN      100     16000 2    998    0.08                               "        AN       100     16000 2    998    0.08                               0.20%    SA       200     1600  4    1996   1.27                               "        ECOLI    100     800   4    1996   2.54                               "        PSA      1200    1600  4    1996   1.25                               "        PC       1800    1600  4    1996   1.25                               "        CAN      100     16000 4    1996   0.16                               "        AN       100     16000 4    1996   0.16                               0.40%    SA       200     1600  8    3992   2.54                               "        ECOLI    100     800   8    3992   5.07                               "        PSA      1200    1600  8    3992   2.50                               "        PC       1800    1600  8    3992   2.50                               "        CAN      100     16000 8    3992   0.33                               "        AN       100     16000 8    3992   0.33                               0.50%    SA       200     1600  10   4900   3.11                               "        ECOLI    100     800   10   4900   6.23                               "        PSA      1200    1600  10   4900   3.07                               "        PC       1800    1600  10   4900   3.07                               "        CAN      100     16000 10   4900   0.41                               "        AN       100     16000 10   4900   0.41                               0.20%    SA       200     1600  4    1996   1.27                               "        ECOLI    100     800   4    1996   2.54                               "        PSA      1200    1600  4    1996   1.25                               "        PC       1800    1600  4    1996   1.25                               "        CAN      100     16000 4    1996   0.16                               "        AN       100     16000 4    1996   0.16                               0.40%    SA       200     1600  8    3992   2.54                               "        ECOLI    100     800   8    3992   5.07                               "        PSA      1200    1600  8    3992   2.50                               "        PC       1800    1600  8    3992   2.50                               "        CAN      100     16000 8    3992   0.33                               "        AN       100     16000 8    3992   0.33                               0.50%    SA       200     1600  10   4900   3.11                               "        ECOLI    100     800   10   4900   6.23                               "        PSA      1200    1600  10   4900   3.07                               "        PC       1800    1600  10   4900   3.07                               "        CAN      100     16000 10   4900   0.41                               "        AN       100     16000 10   4900   0.41                               ______________________________________                                    

                  TABLE 11                                                         ______________________________________                                         200:1 Wt. Ratio DMDMH/IPBC                                                     Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  0.5  99.5   0.06                               "        ECOLI    100     800   0.5  99.5   0.13                               "        PSA      1200    1600  0.5  99.5   0.06                               "        PC       1800    1600  0.5  99.5   0.06                               "        CAN      100     16000 0.5  99.5   0.01                               "        AN       100     16000 0.5  99.5   0.01                                0.025%  SA       200     1600  1.25 248.75 0.16                               "        ECOLI    100     800   1.25 248.75 0.32                               "        PSA      1200    1600  1.25 248.75 0.16                               "        PC       1800    1600  1.25 248.75 0.16                               "        CAN      100     16000 1.25 248.75 0.03                               "        AN       100     16000 1.25 248.75 0.03                               0.05%    SA       200     1600  2.5  497.5  0.32                               "        ECOLI    100     800   2.5  497.5  0.65                               "        PSA      1200    1600  2.5  497.5  0.31                               "        PC       1800    1600  2.5  497.5  0.31                               "        CAN      100     16000 2.5  497.5  0.06                               "        AN       100     16000 2.5  497.5  0.06                               0.10%    SA       200     1600  5    995    0.65                               "        ECOLI    100     800   5    995    1.29                               "        PSA      1200    1600  5    995    0.63                               "        PC       1800    1600  5    995    0.62                               "        CAN      100     16000 5    995    0.11                               "        AN       100     16000 5    995    0.11                               0.20%    SA       200     1600  10   1990   1.29                               "        ECOLI    100     800   10   1990   2.59                               "        PSA      1200    1600  10   1990   1.25                               "        PC       1800    1600  10   1990   1.25                               "        CAN      100     16000 10   1990   0.22                               "        AN       100     16000 10   1990   0.22                               0.40%    SA       200     1600  20   3980   2.59                               "        ECOLI    100     800   20   3980   5.18                               "        PSA      1200    1600  20   3980   2.50                               "        PC       1800    1600  20   3980   2.50                               "        CAN      100     16000 20   3980   0.45                               "        AN       100     16000 20   3980   0.45                               0.50%    SA       200     1600  25   4975   3.23                               "        ECOLI    100     800   25   4975   6.47                               "        PSA      1200    1600  25   4975   3.13                               "        PC       1800    1600  25   4975   3.12                               "        CAN      100     16000 25   4975   0.56                               "        AN       100     16000 25   4975   0.56                               ______________________________________                                    

                  TABLE 12                                                         ______________________________________                                         100:1 Wt. Ratio DMDMH/IPBC                                                     Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  1    99     0.07                               "        ECOLI    100     800   1    99     0.13                               "        PSA      1200    1600  1    99     0.06                               "        PC       1800    1600  1    99     0.06                               "        CAN      100     16000 1    99     0.02                               "        AN       100     16000 1    99     0.02                                0.025%  SA       200     1600  2.5  248    0.17                               "        ECOLI    100     800   2.5  248    0.33                               "        PSA      1200    1600  2.5  248    0.16                               "        PC       1800    1600  2.5  248    0.16                               "        CAN      100     16000 2.5  248    0.04                               "        AN       100     16000 2.5  248    0.04                               0.05%    SA       200     1600  5    495    0.33                               "        ECOLI    100     800   5    495    0.67                               "        PSA      1200    1600  5    495    0.31                               "        PC       1800    1600  5    495    0.31                               "        CAN      100     16000 5    495    0.08                               "        AN       100     16000 5    495    0.08                               0.10%    SA       200     1600  10   990    0.67                               "        ECOLI    100     800   10   990    1.34                               "        PSA      1200    1600  10   990    0.63                               "        PC       1800    1600  10   990    0.62                               "        CAN      100     16000 10   990    0.16                               "        AN       100     16000 10   990    0.16                               0.20%    SA       200     1600  20   1980   1.34                               "        ECOLI    100     800   20   1980   2.68                               "        PSA      1200    1600  20   1980   1.25                               "        PC       1800    1600  20   1980   1.25                               "        CAN      100     16000 20   1980   0.32                               "        AN       100     16000 20   1980   0.32                               0.40%    SA       200     1600  40   3960   2.68                               "        ECOLI    100     800   40   3960   5.35                               "        PSA      1200    1600  40   3960   2.51                               "        PC       1800    1600  40   3960   2.50                               "        CAN      100     16000 40   3960   0.65                               "        AN       100     16000 40   3960   0.65                               0.50%    SA       200     1600  50   4950   3.34                               "        ECOLI    100     800   50   4950   6.69                               "        PSA      1200    1600  50   4950   3.14                               "        PC       1800    1600  50   4950   3.12                               "        CAN      100     16000 50   4950   0.81                               "        AN       100     16000 50   4950   0.81                               ______________________________________                                    

                  TABLE 13                                                         ______________________________________                                         50:1 Wt. Ratio DMDMH/IPBC                                                      Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  2    98     0.07                               "        ECOLI    100     800   2    98     0.14                               "        PSA      1200    1600  2    98     0.06                               "        PC       1800    1600  2    98     0.06                               "        CAN      100     16000 2    98     0.03                               "        AN       100     16000 2    98     0.03                                0.025%  SA       200     1600  5    245    0.18                               "        ECOLI    100     800   5    245    0.36                               "        PSA      1200    1600  5    245    0.16                               "        PC       1800    1600  5    245    0.16                               "        CAN      100     16000 5    245    0.07                               "        AN       100     16000 5    245    0.07                               0.05%    SA       200     1600  10   490    0.36                               "        ECOLI    100     800   10   490    0.71                               "        PSA      1200    1600  10   490    0.31                               "        PC       1800    1600  10   490    0.31                               "        CAN      100     16000 10   490    0.13                               "        AN       100     16000 10   490    0.13                               0.10%    SA       200     1600  20   980    0.71                               "        ECOLI    100     800   20   980    1.43                               "        PSA      1200    1600  20   980    0.63                               "        PC       1800    1600  20   980    0.62                               "        CAN      100     16000 20   980    0.26                               "        AN       100     16000 20   980    0.26                               0.20%    SA       200     1600  40   1960   1.43                               "        ECOLI    100     800   40   1960   2.85                               "        PSA      1200    1600  40   1960   1.26                               "        PC       1800    1600  40   1960   1.25                               "        CAN      100     16000 40   1960   0.52                               "        AN       100     16000 40   1960   0.52                               0.40%    SA       200     1600  80   3920   2.85                               "        ECOLI    100     800   80   3920   5.70                               "        PSA      1200    1600  80   3920   2.52                               "        PC       1800    1600  80   3920   2.49                               "        CAN      100     16000 80   3920   1.05                               "        AN       100     16000 80   3920   1.05                               0.50%    SA       200     1600  100  4900   3.56                               "        ECOLI    100     800   100  4900   7.13                               "        PSA      1200    1600  100  4900   3.15                               "        PC       1800    1600  100  4900   3.12                               "        CAN      100     16000 100  4900   1.31                               "        AN       100     16000 100  4900   1.31                               ______________________________________                                    

                  TABLE 14                                                         ______________________________________                                         20:1 Wt. Ratio DMDMH/IPBC                                                      Use Level                                                                               Organism Q.sub.a Q.sub.b                                                                              Q.sub.A                                                                             Q.sub.B                                                                               SI                                 ______________________________________                                         0.01%    SA       200     1600  5    95     0.08                               "        ECOLI    100     800   5    95     0.17                               "        PSA      1200    1600  5    95     0.06                               "        PC       1800    1600  5    95     0.06                               "        CAN      100     16000 5    95     0.06                               "        AN       100     16000 5    95     0.06                                0.025%  SA       200     1600  12.5 237.5  0.21                               "        ECOLI    100     800   12.5 237.5  0.42                               "        PSA      1200    1600  12.5 237.5  0.16                               "        PC       1800    1600  12.5 237.5  0.16                               "        CAN      100     16000 12.5 237.5  0.14                               "        AN       100     16000 12.5 237.5  0.14                               0.05%    SA       200     1600  25   475    0.42                               "        ECOLI    100     800   25   475    0.84                               "        PSA      1200    1600  25   475    0.32                               "        PC       1800    1600  25   475    0.31                               "        CAN      100     16000 25   475    0.28                               "        AN       100     16000 25   475    0.28                               0.10%    SA       200     1600  50   950    0.84                               "        ECOLI    100     800   50   950    1.69                               "        PSA      1200    1600  50   950    0.64                               "        PC       1800    1600  50   950    0.62                               "        CAN      100     16000 50   950    0.56                               "        AN       100     16000 50   950    0.56                               0.20%    SA       200     1600  100  1900   1.69                               "        ECOLI    100     800   100  1900   3.38                               "        PSA      1200    1600  100  1900   1.27                               "        PC       1800    1600  100  1900   1.24                               "        CAN      100     16000 100  1900   1.12                               "        AN       100     16000 100  1900   1.12                               0.40%    SA       200     1600  200  4800   4.00                               "        ECOLI    100     800   200  4800   8.00                               "        PSA      1200    1600  200  4800   3.17                               "        PC       1800    1600  200  4800   3.11                               "        CAN      100     16000 200  4800   2.30                               "        AN       100     16000 200  4800   2.30                               0.50%    SA       200     1600  250  4750   4.22                               "        ECOLI    100     800   250  4750   8.44                               "        PSA      1200    1600  250  4750   3.18                               "        PC       1800    1600  250  4750   3.11                               "        CAN      100     16000 250  4750   2.80                               "        AN       100     16000 250  4750   2.80                               ______________________________________                                    

Similar SI results also were found with GADM and SUTTOCIDE® A as the methylol compound in place of Germall® II or DMDMH in admixtures with IPBC over the same wt. ratios and use level ranges as shown in the Tables 1-14 above.

Tables 1 through 14 above illustrate the synergism of IPBC (compound B) with Germall® II or DMDMH (compound A) at weight ratios of A:B of 2000:1, 1000:1, 500:1, 200:1, 100:1, 50:1 and 20:1. Synergism is very effective for all ratios at low use levels, e.g. 0.01% to 0.1%, against all tested gram-positive, gram-negative organisms and fungi organisms. At slightly higher use concentrations, e.g. 0.20 to 0.50%, all tested ratios were synergistic against Candida albicans and A. niger also. However, at 50:1 and 20:1 ratios, the synergistic effect is negligible at the 0.01 to 0.1% use levels, and non-synergistic even against Candida albicans and A. niger at use levels of 0.20 to 0.50%.

The SI values were lower for Germall® II as the methylol compound in the admixtures as compared to DMDMH.

Similar results were obtained when iodopropynyl alcohol (IPGA) was substituted for IPBC in the admixtures described above.

3. PRESERVATIVE ACTIVITY (CHALLENGE TEST)

A typical cosmetic emulsion was prepared for microbiological challenge testing and predetermined admixtures of a methylol compound and IPBC were added at various use levels. The emulsion thus prepared had the following composition:

    ______________________________________                                                                   % wt.                                                ______________________________________                                         Phase A                                                                        Stearic Acid                5.00                                               Mineral Oil                 2.50                                               Cetyl Alcohol               1.00                                               Lareth-5 and Ceteth-5 and Oleth-5 and Steareth-5                                                           0.50                                               Glycerol Monostearate and Polyoxyethylene Stearate                                                         1.50                                               Phase B                                                                        Deionized Water             88.0                                               Triethanolamine 99%         1.00                                               Citric Acid 30% aqueous solution                                                                           0.60                                               Preservative Admixture      qs                                                 ______________________________________                                    

To prepare the emulsion, Phases A and B were heated separately to 75°-80° C. Phase A then was added to Phase B with mixing. The mixture then was cooled to 55°-60° C. At this point the desired amount of the preservative admixture was added and the product was cooled to 50° C. while stirring. The citric acid solution then was added to adjust the pH and the mixture was stirred until a temperature of 30° C. was reached.

The challenge tests were carried out using the following microorganisms: SA, ECOLI, PSA, PC, AN and CAN, in this manner. 50 g. aliquots of the test emulsion containing various amounts of the preservative admixture were inoculated with approximately 10⁷ -10⁸ of the challenge organisms. The test samples then were stirred to disperse the challenge inoculum. The samples were incubated and assayed at 48 hours, 7, 14, 21 and 28 days. The assays were performed on 1 g. of the test sample by serially diluting 10¹ to 10⁶ of the original concentration. The plating medium for bacteria was Letheen agar and for fungi it was low pH Mycophil agar with Tween 20. Each plated sample was incubated for 48 hours at 37° C. for bacteria, 5 days at 25° C. for mold, and 3 days at 25° C. for fungi. After incubation, readings of the number of colonies per milliliter (cfu/ml) were made. At 21 days the test product was reinoculated with half of the original inoculum. The data is presented in Tables 15-23 below.

                  TABLE 15                                                         ______________________________________                                         2000:1 GERMALL ® II/IPBC                                                                    48      7     14    21    28                                  Test Level                                                                             Organism Hours   Days  Days  Days  Days                                ______________________________________                                          0.01%  AN       69,000  260,000                                                                              190,000                                                                              17,000                                                                               4,500                               "       CAN      98,000  76,000                                                                               1,400 3,100 19,000                              "       ECOLI    110,000 290,000                                                                              2,400 138,000                                                                              560,000                             "       PC       <10     <10   <10   <10   <10                                 "       PSA      <10     <10   <10   <10   <10                                 "       SA       190,000 220   <10   <10   3,700                                0.025% AN       2,800   10    10    <10   220                                 "       CAN      58,000  29,000                                                                               18,000                                                                               56,000                                                                               110,000                             "       ECOLI    39,000  10    <10   <10   <10                                 "       PC       <10     <10   <10   <10   <10                                 "       PSA      <10     <10   <10   <10   <10                                 "       SA       37,000  170   <10   <10   <10                                  0.05%  AN       20      <10   <10   <10   <10                                 "       CAN      19,000  6,600 70    <10   320                                 "       ECOLI    3,400   <10   <10   <10   <10                                 "       PC       <10     <10   <10   <10   <10                                 "       PSA      <10     <10   <10   <10   <10                                 "       SA       31,000  <10   <10   <10   <10                                 0.1%    AN       <10     <10   <10   <10   <10                                 "       CAN      180     <10   <10   <10   <10                                 "       ECOLI    <10     <10   <10   <10   <10                                 "       PC       <10     <10   <10   <10   <10                                 "       PSA      <10     <10   <10   <10   <10                                 "       SA       750     <10   <10   <10   <10                                 0.2%    AN       <10     <10   <10   <10   <10                                 "       CAN      <10     <10   <10   <10   <10                                 "       ECOLI    <10     <10   <10   <10   <10                                 "       PC       <10     <10   <10   <10   <10                                 "       PSA      <10     <10   <10   <10   <10                                 "       SA       <10     <10   <10   <10   <10                                 ______________________________________                                         Unpreserved control                                                            Organism                                                                               48 Hours 7 Days   14 Days                                                                               21 Days                                                                               28 Days                                ______________________________________                                         AN      52,000   27,000   19,000 19,000 19,000                                 CAN     110,000  130,000  240,000                                                                               180,000                                                                               240,000                                ECOLI   54,000   140,000  170,000                                                                               170,000                                                                               74,000                                 PC      6,400,000                                                                               6,400,000                                                                               2,000,000                                                                             6,700,000                                                                             29,000                                 PSA     110,000  700      110,000                                                                               290,000                                                                               85,000                                 SA      2,800,000                                                                               250,000  51,000 3,700  330                                    ______________________________________                                         Inoculum Concentration                                                         Organism       0 Hours  21 Days                                                ______________________________________                                         AN             26,000   53,000                                                 CAN            1,000,000                                                                               1,900,000                                              ECOLI          3,600,000                                                                               170,000                                                PC             3,400,000                                                                               87,000                                                 PSA            4,500,000                                                                               390,000                                                SA             4,100,000                                                                               200,000                                                ______________________________________                                    

                  TABLE 16                                                         ______________________________________                                         1000:1 GERMALL ® II/IPBC                                                   Test           48      7     14    21    28                                    Level Organism Hours   Days  Days  Days  Days                                  ______________________________________                                         0.01% AN       34,000  3,500 80    10    <10                                   "     CAN      420,000 24,000                                                                               950   6,400 6,400,000                             "     ECOLI    120,000 63,000                                                                               93,000                                                                               92,000                                                                               9,600,000                             "     PC       10      25,000                                                                               1,600 15,800                                                                               33,000,000                            "     PSA      <10     <10   <10   <10   <10                                   "     SA       100,000 1,400 <10   <10   5,000                                 0.025%                                                                               AN       530     10    <10   <10   <10                                   "     CAN      34,000  750   10    770   240,000                               "     ECOLI    120,000 <10   <10   <10   <10                                   "     PC       <10     <10   <10   <10   <10                                   "     PSA      <10     <10   <10   <10   <10                                   "     SA       37,000  170   <10   <10   <10                                   0.05% AN       <10     <10   <10   <10   <10                                   "     CAN      13,000  <10   <10   <10   <10                                   "     ECOLI    68,000  <10   <10   <10   <10                                   "     PC       <10     <10   <10   <10   <10                                   "     PSA      <10     <10   <10   <10   <10                                   "     SA       21,000  <10   <10   <10   <10                                   0.1%  AN       <10     <10   <10   <10   <10                                   "     CAN      10      <10   <10   <10   <10                                   "     ECOLI    <10     <10   <10   <10   <10                                   "     PC       <10     <10   <10   <10   <10                                   "     PSA      <10     <10   <10   <10   <10                                   "     SA       1,400   <10   <10   <10   <10                                   0.2%  AN       <10     <10   <10   <10   <10                                   "     CAN      <10     <10   <10   <10   <10                                   "     ECOLI    <10     <10   <10   <10   <10                                   "     PC       <10     <10   <10   <10   <10                                   "     PSA      <10     <10   <10   <10   <10                                   "     SA       <10     <10   <10   <10   <10                                   ______________________________________                                         Unpreserved control                                                            Organism                                                                               48 Hours 7 Days   14 Days                                                                               21 Days                                                                               28 Days                                ______________________________________                                         AN      52,000   27,000   19,000 19,000 19,000                                 CAN     110,000  130,000  240,000                                                                               180,000                                                                               240,000                                ECOLI   54,000   140,000  170,000                                                                               170,000                                                                               74,000                                 PC      6,400,000                                                                               6,400,000                                                                               2,000,000                                                                             6,700,000                                                                             29,000                                 PSA     110,000  700      110,000                                                                               290,000                                                                               85,000                                 SA      2,800,000                                                                               250,000  51,000 3,700  330                                    ______________________________________                                         Inoculum Concentration                                                         Organism       0 Hours  21 Days                                                ______________________________________                                         AN             26,000   53,000                                                 CAN            1,000,000                                                                               1,900,000                                              ECOLI          3,600,000                                                                               170,000                                                PC             3,400,000                                                                               87,000                                                 PSA            4,500,000                                                                               390,000                                                SA             4,100,000                                                                               200,000                                                ______________________________________                                    

                                      TABLE 17                                     __________________________________________________________________________     500:1 GERMALL ® II/IPBC                                                    Test Level                                                                           Organism                                                                             48 Hours                                                                            7 Days                                                                              14 Days                                                                             21 Days                                                                              28 Days                                       __________________________________________________________________________      0.01%                                                                               AN    23,000                                                                              40   <10  <10   <10                                           "     CAN   170,000                                                                             5,600                                                                               290  200   <10                                           "     ECOLI 90,000                                                                              57,000                                                                              95,000                                                                              70,000                                                                               240,000                                       "     PC    10   <10  <10  >10,000                                                                              42,000,000                                    "     PSA   <10  <10  <10  <10   <10                                           "     SA    380,000                                                                             440  <10  <10   5,100                                          0.05%                                                                               AN    <10  <10  <10  <10   <10                                           "     CAN   8,700                                                                               <10  <10  <10   <10                                           "     ECOLI 60,000                                                                              <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    31,000                                                                              <10  <10  <10   <10                                           0.1%  AN    <10  <10  <10  <10   <10                                           "     CAN   <10  <10  <10  <10   <10                                           "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    890  <10  <10  <10   <10                                           0.2%  AN    <10  <10  <10  <10   <10                                           "     CAN   <10  <10  <10  <10   <10                                           "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    <10  <10  <10  <10   <10                                           __________________________________________________________________________     Unpreserved control                                                            Organism                                                                            48 Hours                                                                               7 Days                                                                               14 Days                                                                               21 Days                                                                             28 Days                                         __________________________________________________________________________     AN   6,100   520,000                                                                              18,000 5,000                                                                               11,000                                          CAN  1,000,000                                                                              710,000                                                                              95,000 12,000                                                                              64,000                                          ECOLI                                                                               7,100,000                                                                              6,200,000                                                                            610,000                                                                               350,000                                                                             120,000                                         PC   14,600,000                                                                             160,000,000                                                                          3,600,000                                                                             2,720,000                                                                           9,500,000                                       PSA  20      900   130    4,100                                                                               >100,000                                        SA   43,000,000                                                                             600,000                                                                              1,000  220  <10                                             __________________________________________________________________________     Inoculum Concentration                                                         Organism     0 Hours                                                                             21 Days                                                      __________________________________________________________________________     AN           19,000                                                                              4,700,000                                                    CAN          340,000                                                                             16,000,000                                                   ECOLI        3,900,000                                                                           1,480,000                                                    PC           3,800,000                                                                           1,380,000                                                    PSA          9,200,000                                                                           730,000                                                      SA           4,800,000                                                                           360,000                                                      __________________________________________________________________________

                                      TABLE 18                                     __________________________________________________________________________     200:1 GERMALL ® II/IPBC                                                    Test Level                                                                           Organism                                                                             48 Hours                                                                            7 Days                                                                              14 Days                                                                             21 Days                                                                              28 Days                                       __________________________________________________________________________      0.01%                                                                               AN    47,000                                                                              320  10   <10   <10                                           "     CAN   810,000                                                                             450,000                                                                             410,000                                                                             190,000                                                                              63,000                                        "     ECOLI 220,000                                                                             7,600                                                                               <10  850   >1,000,000                                    "     PC    10,000                                                                              500,000                                                                             1,900,000                                                                           1,100,000                                                                            193,000                                       "     PSA   <10  <10  <10  <10   <10                                           "     SA    190,000                                                                             23,000                                                                              120  <10   <10                                            0.05%                                                                               AN    <10  <10  <10  <10   <10                                           "     CAN   190  <10  <10  <10   <10                                           "     ECOLI 37,000                                                                              <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    19,000                                                                              <10  <10  <10   <10                                           0.1%  AN    <10  <10  <10  <10   <10                                           "     CAN   10   <10  <10  <10   <10                                           "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    45,000                                                                              <10  <10  <10   <10                                           0.2%  AN    <10  <10  <10  <10   <10                                           "     CAN   <10  <10  <10  <10   <10                                           "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    <10  <10  <10  <10   <10                                           __________________________________________________________________________     Unpreserved control                                                            Organism                                                                            48 Hours                                                                               7 Days                                                                               14 Days                                                                               21 Days                                                                             28 Days                                         __________________________________________________________________________     AN   89,000  32,000                                                                               22,000 16,000                                                                              16,000                                          CAN  210,000 670,000                                                                              430,000                                                                               590,000                                                                             640,000                                         ECOLI                                                                               640,000 360,000                                                                              410,000                                                                               990,000                                                                             68,000                                          PC   19,000,000                                                                             3,200,000                                                                            7,000,000                                                                             >10,000                                                                             2,760,000                                       PSA  80      9,400 200,000                                                                               >10,000                                                                             34,000                                          SA   6,300,000                                                                              190,000                                                                              11,000 580  120                                             __________________________________________________________________________     Inoculum Concentration                                                         Organism     0 Hours                                                                             21 Days                                                      __________________________________________________________________________     AN           41,000                                                                              32,000                                                       CAN          640  1,100,000                                                    ECOLI        5,800,000                                                                           1,300,000                                                    PC           900,000                                                                             3,000,000                                                    PSA          1,800,000                                                                           4,900,000                                                    SA           7,200,000                                                                           2,000,000                                                    __________________________________________________________________________

                  TABLE 19                                                         ______________________________________                                         50:1 GERMALL ® II/IPBC                                                                      48      7     14    21    28                                  Test Level                                                                             Organism Hours   Days  Days  Days  Days                                ______________________________________                                          0.01%  AN       4,100   40    <10   <10   <10                                 "       CAN      310,000 7,700 560   5,600 5,200                               "       ECOLI    170,000 710   10    <10   120                                 "       PC       7,400   74,000                                                                               340,000                                                                              720,000                                                                              520,000                             "       PSA      <10     <10   <10   <10   30                                  "       SA       110,000 11,000                                                                               <10   <10   9,200                                0.05%  AN       <10     100   <10   <10   <10                                 "       CAN      210     <10   <10   <10   <10                                 "       ECOLI    150,000 <10   <10   <10   <10                                 "       PC       <10     <10   <10   <10   <10                                 "       PSA      <10     <10   <10   <10   <10                                 "       SA       35,000  <10   <10   <10   <10                                 0.1%    AN       <10     <10   <10   <10   <10                                 "       CAN      <10     <10   <10   <10   <10                                 "       ECOLI    510     <10   <10   <10   <10                                 "       PC       <10     <10   <10   <10   <10                                 "       PSA      <10     <10   <10   <10   <10                                 "       SA       3,000   <10   <10   <10   <10                                 0.2%    AN       <10     <10   <10   <10   <10                                 "       CAN      <10     <10   <10   <10   <10                                 "       ECOLI    <10     <10   <10   <10   <10                                 "       PC       <10     <10   <10   <10   <10                                 "       PSA      <10     <10   <10   <10   <10                                 "       SA       <10     <10   <10   <10   <10                                 ______________________________________                                         Unpreserved control                                                            Organism                                                                              48 Hours  7 Days   14 Days                                                                               21 Days                                                                               28 Days                                ______________________________________                                         AN     89,000    32,000   22,000 16,000 16,000                                 CAN    210,000   670,000  430,000                                                                               590,000                                                                               640,000                                ECOLI  640,000   360,000  410,000                                                                               990,000                                                                               68,000                                 PC     19,000,000                                                                               3,200,000                                                                               7,000,000                                                                             >10,000                                                                               2,760,000                              PSA    80        9,400    200,000                                                                               >10,000                                                                               34,000                                 SA     6,300,000 190,000  11,000 580    120                                    ______________________________________                                         Inoculum Concentration                                                         Organism       0 Hours  21 Days                                                ______________________________________                                         AN             41,000   32,000                                                 CAN            640      1,100,000                                              ECOLI          5,800,000                                                                               1,300,000                                              PC             900,000  3,000,000                                              PSA            1,800,000                                                                               4,900,000                                              SA             7,200,000                                                                               2,000,000                                              ______________________________________                                    

                                      TABLE 20                                     __________________________________________________________________________     20:1 GERMALL ® II/IPBC                                                     Test Level                                                                           Organism                                                                             48 Hours                                                                            7 Days                                                                              14 Days                                                                             21 Days                                                                              28 Days                                       __________________________________________________________________________      0.01%                                                                               AN    3,100                                                                               <10  <10  <10   <10                                           "     CAN   75,000                                                                              220  <10  <10   2,400                                         "     ECOLI 160,000                                                                             110  <10  <10   20                                            "     PC    12,000                                                                              1,000,000                                                                           2,100,000                                                                           >1,000,000                                                                           730,000                                       "     PSA   <10  <10  <10  <10   4,000                                         "     SA    140,000                                                                             4,100                                                                               <10  <10   1,680                                          0.05%                                                                               AN    <10  <10  <10  <10   <10                                           "     CAN   <10  <10  <10  <10   <10                                           "     ECOLI 16,000                                                                              <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10                                                 "     SA    31,000                                                                              <10  <10  <10   <10                                           0.1%  AN    <10  <10  <10  <10   <10                                           "     CAN   <10  <10  <10  <10   <10                                           "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    6,800                                                                               <10  <10  <10   <10                                           0.2%  AN    <10  <10  <10  <10   <10                                           "     CAN   <10  <10  <10  <10   <10                                           "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    <10  <10  <10  <10   <10                                           __________________________________________________________________________     Unpreserved control                                                            Organism                                                                            48 Hours                                                                               7 Days                                                                               14 Days                                                                               21 Days                                                                             28 Days                                         __________________________________________________________________________     AN   89,000  32,000                                                                               22,000 16,000                                                                              16,000                                          CAN  210,000 670,000                                                                              430,000                                                                               590,000                                                                             640,000                                         ECOLI                                                                               640,000 360,000                                                                              410,000                                                                               990,000                                                                             68,000                                          PC   19,000,000                                                                             3,200,000                                                                            7,000,000                                                                             >10,000                                                                             2,760,000                                       PSA  80      9,400 200,000                                                                               >10,000                                                                             34,000                                          SA   6,300,000                                                                              190,000                                                                              11,000 580  120                                             __________________________________________________________________________     Inoculum Concentration                                                         Organism     0 Hours                                                                             21 Days                                                      __________________________________________________________________________     AN           41,000                                                                              32,000                                                       CAN          640  1,100,000                                                    ECOLI        5,800,000                                                                           1,300,000                                                    PC           900,000                                                                             3,000,000                                                    PSA          1,800,000                                                                           4,900,000                                                    SA           7,200,000                                                                           2,000,000                                                    __________________________________________________________________________

                                      TABLE 21                                     __________________________________________________________________________     2000:1 DMDMH/IPBC                                                              Test Level                                                                           Organism                                                                             48 Hours                                                                            7 Days                                                                              14 Days                                                                             21 Days                                                                              28 Days                                       __________________________________________________________________________      0.025%                                                                              AN    4,100                                                                               38,000                                                                              270  80    100                                           "     CAN   270,000                                                                             1,900,000                                                                           550,000                                                                             220,000                                                                              210,000                                       "     ECOLI 1,300,000                                                                           <10  <10  <10   3,000                                         "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    42,000                                                                              20   <10  <10   <10                                            0.05%                                                                               AN    <10  <10  <10  <10   <10                                           "     CAN   770,000                                                                             130,000                                                                             670,000                                                                             160,000                                                                              64,000                                        "     ECOLI 220,000                                                                             <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    9,000                                                                               <10  <10  <10   <10                                           0.1%  AN    <10  <10  <10  <10   <10                                           "     CAN   580  840  500  40,000                                                                               83,000                                        "     ECOLI 340  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    29,000                                                                              <10  <10  <10   <10                                           0.2%  AN    <10  <10  <10  <10   <10                                           "     CAN   <10  10   10   <10   21                                            "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    <10  <10  <10  <10   <10                                           __________________________________________________________________________     Unpreserved control                                                            Organism                                                                            48 Hours                                                                               7 Days                                                                               14 Days                                                                               21 Days                                                                             28 Days                                         __________________________________________________________________________     AN   37,000  36,000                                                                               24,000 5,200                                                                               6,000                                           CAN  120,000 1,900,000                                                                            3,300,000                                                                             480,000                                                                             790,000                                         ECOLI                                                                               150,000 2,500,000                                                                            7,300,000                                                                             240,000                                                                             140,000                                         PC   19,000,000                                                                             15,600,000                                                                           5,900,000                                                                             8,500,000                                                                           31,000,000                                      PSA  <10     <10   100    15,200                                                                              300,000                                         SA   7,000,000                                                                              >1,000,000                                                                           12,000 3,000                                                                               110                                             __________________________________________________________________________     Inoculum Concentration                                                         Organism     0 Hours                                                                             21 Days                                                      __________________________________________________________________________     AN           50,000                                                                              41,000                                                       CAN          1,400,000                                                                           640                                                          ECOLI        4,800,000                                                                           5,800,000                                                    PC           9,200,000                                                                           900,000                                                      PSA          6,900,000                                                                           9,000,000                                                    SA           5,700,000                                                                           7,200,000                                                    __________________________________________________________________________

                                      TABLE 22                                     __________________________________________________________________________     1000:1 DMDMH/IPBC                                                              Test Level                                                                           Organism                                                                             48 Hours                                                                            7 Days                                                                              14 Days                                                                             21 Days                                                                              28 Days                                       __________________________________________________________________________      0.025%                                                                              AN    650  900  <10  <10   140                                           "     CAN   97,000                                                                              900,000                                                                             2,300,000                                                                           120,000                                                                              120,000                                       "     ECOLI 160,000                                                                             <10  <10  <10   2,400                                         "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    23,000                                                                              <10  <10  <10   90                                             0.05%                                                                               AN    20   <10  <10  <10   <10                                           "     CAN   65,000                                                                              520,000                                                                             56,000                                                                              32,000                                                                               370,000                                       "     ECOLI 26,000                                                                              <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    12,000                                                                              <10  <10  <10   <10                                           0.1%  AN    <10  <10  <10  <10   <10                                           "     CAN   3,100                                                                               4,800                                                                               180  640   4,400                                         "     ECOLI 45,000                                                                              <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    6,000                                                                               <10  <10  <10   <10                                           0.2%  AN    <10  <10  <10  <10   <10                                           "     CAN   <10  <10  <10  <10   <10                                           "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    180  <10  <10  <10   <10                                           __________________________________________________________________________     Unpreserved control                                                            Organism                                                                            48 Hours                                                                               7 Days                                                                               14 Days                                                                               21 Days                                                                             28 Days                                         __________________________________________________________________________     AN   37,000  36,000                                                                               24,000 5,200                                                                               6,000                                           CAN  120,000 1,900,000                                                                            3,300,000                                                                             480,000                                                                             790,000                                         ECOLI                                                                               150,000 2,500,000                                                                            7,300,000                                                                             240,000                                                                             140,000                                         PC   19,000,000                                                                             15,600,000                                                                           5,900,000                                                                             8,500,000                                                                           31,000,000                                      PSA  <10     <10   100    15,200                                                                              300,000                                         SA   7,000,000                                                                              >1,000,000                                                                           12,000 3,000                                                                               110                                             __________________________________________________________________________     Inoculum Concentration                                                         Organism     0 Hours                                                                             21 Days                                                      __________________________________________________________________________     AN           50,000                                                                              41,000                                                       CAN          1,400,000                                                                           640                                                          ECOLI        4,800,000                                                                           5,800,000                                                    PC           9,200,000                                                                           900,000                                                      PSA          6,900,000                                                                           9,000,000                                                    SA           5,700,000                                                                           7,200,000                                                    __________________________________________________________________________

                                      TABLE 23                                     __________________________________________________________________________     2000:1 GADM/IPBC                                                               Test Level                                                                           Organism                                                                             48 Hours                                                                            7 Days                                                                              14 Days                                                                             21 Days                                                                              28 Days                                       __________________________________________________________________________      0.025%                                                                              AN    300  21   10   <10   11                                            "     CAN   480,000                                                                             890,000                                                                             940,000                                                                             1,040,000                                                                            130,000                                       "     ECOLI 230,000                                                                             <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    78,000                                                                              <10  <10  <10   <10                                            0.05%                                                                               AN    <10  <10  <10  <10   <10                                           "     CAN   110,000                                                                             4,100,000                                                                           3,600,000                                                                           330,000                                                                              97,000                                        "     ECOLI 120,000                                                                             <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    56,000                                                                              <10  <10  <10   <10                                           0.1%  AN    <10  <10  <10  <10   <10                                           "     CAN   1,200,000                                                                           53,000                                                                              430,000                                                                             144,000                                                                              110,000                                       "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    <10  <10  <10  <10   <10                                           0.2%  AN    <10  <10  <10  <10   <10                                           "     CAN   90,000                                                                              <10  <10  <10   60                                            "     ECOLI <10  <10  <10  <10   <10                                           "     PC    <10  <10  <10  <10   <10                                           "     PSA   <10  <10  <10  <10   <10                                           "     SA    <10  <10  <10  <10   <10                                           __________________________________________________________________________     Unpreserved control                                                            Organism                                                                            48 Hours                                                                               7 Days                                                                               14 Days                                                                               21 Days                                                                              28 Days                                        __________________________________________________________________________     AN   50,000  33,000                                                                               33,000 13,000                                                                               5,400                                          CAN  780,000 780,000                                                                              780,000                                                                               500,000                                                                              170,000                                        ECOLI                                                                               600,000 3,100,000                                                                            920,000                                                                               920,000                                                                              140,000                                        PC   11,000,000                                                                             30,000,000                                                                           10,000,000                                                                            10,000,000                                                                           1,400,000                                      PSA  3,800   600   12,800 12,800                                                                               100,000                                        SA   14,000,000                                                                             410,000                                                                              7,100  7,100 80                                             __________________________________________________________________________     Inoculum Concentration                                                         Organism     0 Hours                                                                             21 Days                                                      __________________________________________________________________________     AN           53,000                                                                              10,000                                                       CAN          1,900,000                                                                           310,000                                                      ECOLI        170,000                                                                             3,500,000                                                    PC           87,000                                                                              2,500,000                                                    PSA          390,000                                                                             5,400,000                                                    SA           200,000                                                                             4,100,000                                                    __________________________________________________________________________

Discussion of Challenge Testing Results

The 28-day challenge results reported in Tables 15-23 above demonstrate the effectiveness of the preservative admixture of the invention in a use emulsion composition against a wide range of bacteria and fungi organisms.

For example, admixture compositions of Germall® II and IPBC at a wt. ratio of 2000:1 (Table 15), when present at use levels of 0.05 to 0.2%, corresponding to 0.75 to 10 ppm IPBC and 500 to 2000 ppm methylol levels, provide substantially complete protection against all tested organisms after 28 days. At the low use level of 0.05% active, all the challenge tests passed within 21 days. Then, upon reinoculation after 21 days, all organisms died within 7 days except CAN which cleared within 14 days.

Table 21 shows the challenge test results for DMDMH and IPBC admixtures at the same 2000:1 wt. ratio. A use level of 0.2%, however, is needed for this blend to pass against all organisms after 21 days. Upon reinoculation, all organisms died within 7 days with the exception of CAN which cleared within 14 days. It is thus evident from these results that Germall® II blended with IPBC is 4 times more effective than a DMDMH/IPBC blend.

Germall® II also is superior to GADM as the methylol compound, as shown in Table 23.

Table 24 below is a study of the activity of solution of GII/IPBC in propylene glycol. The admixtures of the active GII and IPBC components were prepared at weight ratios of 99.5%/0.5% and 99%/1%, and added to 60% by weight propylene glycol. The resulting solutions were tested at 0.05%, 0.1% and 0.2% total active in a proteinaceous shampoo formulation.

                                      TABLE 24                                     __________________________________________________________________________     Test Level                                                                           Organism                                                                            48 Hours                                                                            7 Days                                                                               14 Days                                                                             21 Days                                                                               28 Days                                      __________________________________________________________________________     99.5% GII/0.5% IPBC/propylene glycol solution - 0.05% total active              0.05%                                                                               AN   310,000                                                                             2,000 <10  <10    <10                                          "     CAN  44,000                                                                              2,400 <10  <10    1,800                                        "     ECOLI                                                                               3,000                                                                               <10   <10  <10    <10                                          "     PC   400,000                                                                             62,000                                                                               90   <10    >10,000                                      "     PSA  5,300,000                                                                           3,000 <10  <10    >10,000                                      "     SA   10   10    <10  <10    <10                                          99.5% GII/0.5% IPBC/propylene glycol solution - 0.1% total active              0.1%  AN   210,000                                                                             <10   <10  <10    <10                                          "     CAN  1,500                                                                               <10   <10  <10    <10                                          "     ECOLI                                                                               580  <10   <10  <10    20                                           "     PC   34,000                                                                              <10   <10  <10    <10                                          "     PSA  780  <10   <10  <10    <10                                          "     SA   <10  <10   <10  <10    <10                                          99.5% GII/0.5% IPBC/propylene glycol solution - 0.2% total active              0.2%  AN   <10  <10   <10  <10    <10                                          "     CAN  <10  <10   <10  <10    <10                                          "     ECOLI                                                                               <10  <10   <10  <10    <10                                          "     PC   <10  <10   <10  <10    <10                                          "     PSA  <10  <10   <10  <10    <10                                          "     SA   <10  <10   <10  <10    <10                                          99% GII/1% IPBC/propylene glycol solution - 0.05% total active                  0.05%                                                                               AN   89,000                                                                              <10   <10  <10    <10                                          "     CAN  4,400                                                                               <10   <10  <10    20                                           "     ECOLI                                                                               3,300                                                                               <10   <10  <10    60                                           "     PC   260,000                                                                             <10   <10  <10    3,400                                        "     PSA  64,000                                                                              <10   <10  <10    280                                          "     SA   <10  <10   <10  <10    <10                                          99% GII/1% IPBC/propylene glycol solution - 0.1% total active                  0.1%  AN   37,000                                                                              <10   <10  <10    <10                                          "     CAN  <10  <10   <10  <10    <10                                          "     ECOLI                                                                               840  <10   <10  <10    <10                                          "     PC   48,000                                                                              <10   <10  <10    <10                                          "     PSA  440  <10   <10  <10    <10                                          "     SA   <10  <10   <10  <10    <10                                          99% GII/1% IPBC/propylene glycol solution - 0.2% total active                  0.2%  AN   <10  <10   <10  <10    <10                                          "     CAN  <10  <10   <10  <10    <10                                          "     ECOLI                                                                               <10  <10   <10  <10    <10                                          "     PC   <10  <10   <10  <10    <10                                          "     PSA  <10  <10   <10  <10    <10                                          "     SA   <10  <10   <10  <10    <10                                          __________________________________________________________________________     Unpreserved control                                                            Organism                                                                             48 Hours                                                                              7 Days 14 Days                                                                              21 Days                                                                               28 Days                                       __________________________________________________________________________     AN    4,000,000                                                                CAN   160,000                                                                               >100,000,000                                                                          5,600,000                                                                            >100,000,000                                                                          >1,000,000                                    ECOLI        >100,000,000                                                                          800,000,000                                                                          48,000,000                                                                            600,000                                       PC    >100,000,000                                                                          50,000,000                                                                            48,000,000   >1,000,000                                    PSA   >100,000,000                                                                          >100,000,000                                                                          58,000,000                                                                           65,000,000                                                                            >1,000,000                                    SA           >100,000,000                                                      __________________________________________________________________________     Inoculum Concentration                                                         Organism     0 Hours                                                                             21 Days                                                      __________________________________________________________________________     AN           380,000                                                                             330,000                                                      CAN          860,000                                                                             4,200,000                                                    ECOLI        2,500,000                                                                           5,300,000                                                    PC           1,900,000                                                                           60,000,000                                                   PSA          3,200,000                                                                           5,000,000                                                    SA           2,400,000                                                                           3,000,000                                                    __________________________________________________________________________

Table 25 shows a similar study as in Table 24 above in which the vehicle for the composition was the typical emulsion described above.

                                      TABLE 25                                     __________________________________________________________________________     Test Level                                                                           Organism                                                                            48 Hours                                                                            7 Days                                                                               14 Days                                                                             21 Days                                                                               28 Days                                      __________________________________________________________________________     99.5% GII/0.5% IPBC/propylene glycol solution - 0.05% total active              0.05%                                                                               AN   40   <10   <10  <10    <10                                          "     CAN  520,000                                                                             <10   <10  <10    <10                                          "     ECOLI                                                                               320  <10   <10  <10    <10                                          "     PC   <10  <10   <10  <10    <10                                          "     PSA  <10  <10   <10  <10    <10                                          "     SA   24,000                                                                              <10   <10  <10    <10                                          99.5% GII/0.5% IPBC/propylene glycol solution - 0.1% total active              0.1%  AN   <10  <10   <10  <10    <10                                          "     CAN  <10  <10   <10  <10    <10                                          "     ECOLI                                                                               10   <10   <10  <10    <10                                          "     PC   <10  <10   <10  <10    <10                                          "     PSA  <10  <10   <10  <10    <10                                          "     SA   650  <10   <10  <10    <10                                          99.5% GII/0.5% IPBC/propylene glycol solution - 0.2% total active              0.2%  AN   <10  <10   <10  <10    <10                                          "     CAN  <10  <10   <10  <10    <10                                          "     ECOLI                                                                               <10  <10   <10  <10    <10                                          "     PC   <10  <10   <10  <10    <10                                          "     PSA  <10  <10   <10  <10    <10                                          "     SA   <10  <10   <10  <10    <10                                          99% GII/1% IPBC/propylene glycol solution - 0.05% total active                  0.05%                                                                               AN   <10  <10   <10  <10    <10                                          "     CAN  6,700                                                                               <10   <10  <10    <10                                          "     ECOLI                                                                               66,000                                                                              <10   <10  <10    <10                                          "     PC   <10  <10   <10  <10    <10                                          "     PSA  <10  <10   <10  <10    <10                                          "     SA   50,000                                                                              <10   <10  <10    <10                                          99% GII/1% IPBC/propylene glycol solution - 0.1% total active                  0.1%  AN   <10  <10   <10  <10    <10                                          "     CAN  <10  <10   <10  <10    <10                                          "     ECOLI                                                                               <10  <10   <10  <10    <10                                          "     PC   <10  <10   <10  <10    <10                                          "     PSA  <10  <10   <10  <10    <10                                          "     SA   620  <10   <10  <10    <10                                          99% GII/1% IPBC/propylene glycol solution - 0.2% total active                  0.2%  AN   <10  <10   <10  <10    <10                                          "     CAN  <10  <10   <10  <10    <10                                          "     ECOLI                                                                               <10  <10   <10  <10    <10                                          "     PC   <10  <10   <10  <10    <10                                          "     PSA  <10  <10   <10  <10    <10                                          "     SA   <10  <10   <10  <10    <10                                          __________________________________________________________________________     Unpreserved control                                                            Organism                                                                             48 Hours                                                                              7 Days 14 Days                                                                              21 Days                                                                               28 Days                                       __________________________________________________________________________     AN    3,100,000                                                                             650,000                                                                               370,000                                                                              1,400,000                                                                             260,000                                       CAN   6,000,000                                                                             4,000,000                                                                             1,100,000                                                                            5,800,000                                                                             >1,000,000                                    ECOLI 11,000,000                                                                            7,300,000                                                                             6,000,000                                                                            730,000                                                                               220,000                                       PC    100,000,000                                                                           53,000,000                                                                            40,000,000                                                                           40,000,000                                                                            600,000                                       PSA   5,000,000                                                                             500,000                                                                               2,700,000                                                                            72,000 20,000                                        SA    30,000,000                                                                            150,000                                                                               440,000                                                                              2,500  9,900                                         __________________________________________________________________________     Inoculum Concentration                                                         Organism     0 Hours                                                                             21 Days                                                      __________________________________________________________________________     AN           380,000                                                                             330,000                                                      CAN          860,000                                                                             4,200,000                                                    ECOLI        2,500,000                                                                           5,300,000                                                    PC           1,900,000                                                                           60,000,000                                                   PSA          3,200,000                                                                           5,000,000                                                    SA           2,400,000                                                                           3,000,000                                                    __________________________________________________________________________

The results shown in Tables 24 and 25 demonstrate that the compositions of the invention are completely effective against the tested organisms in comparison to the unpreserved controls.

While the invention has been described with particular reference to certain embodiments thereof, it will be understood that changes and modifications may be made which are within the skill of the art. Accordingly, it is intended to be bound only by the following claims, in which: 

What is claimed is:
 1. A water soluble preservative antimicrobial composition for addition to commercial use products at predetermined use levels to provide synergistic biocidal activity against a wide range of fungi and gram-negative and gram-positive bacteria, comprising an active liquid blend of(1) an admixture of(a) 1,3-dimethylol-5,5-dimethyl hydantoin; and (b) 3-iodo-2-propynylbutyl carbamate, in a weight ratio of (a):(b) of 100:1 to 2000:1, and (2) propylene glycol or 1,3-butylene glycol.
 2. A water soluble preservative antimicrobial composition according to claim 1 wherein said weight ratio is 200:1 to 500:1.
 3. A water soluble preservative antimicrobial composition according to claim 1 which comprises about 20 to 55 weight percent of (1) and 45 to 80 weight percent of (2).
 4. A composition according to claim 3 which comprises about 40 weight percent of (1) and about 60 weight percent of (2).
 5. A commercial use product which is protected for an extended period of time against contamination by a wide range of fungi and gram-negative and gram-positive bacteria which includes 0.1 to 5% by weight of the composition of claim
 1. 6. A product according to claim 5 which includes about 0.5 to 1% by weight of the composition of claim
 1. 7. A commercial use product according to claim 5 which is a personal care, household or industrial composition.
 8. A commercial use product according to claim 5 which is a shampoo. 